Indigenous Soil Bacterial Communities Research Articles

Co-exposure to pentachlorophenol (PCP) and cadmium (Cd) triggers apoptosis-like cell death in Eschericia coli

Pentachlorophenol (PCP) - cadmium (Cd) complex pollution has been identified as a form of persistent soil pollution in south China, exerting detrimental impacts on the indigenous soil bacterial communities. Hence, it is worthwhile to investigate whether and how bacterial populations alter in response to these pollutants. In this study, Escherichia coli was used as a model bacterium. Results showed that PCP exposure caused bacterial cell membrane permeability changes, intracellular ROS elevation, and DNA fragmentation, and triggered apoptosis-like cell death at low exposure concentration and necrosis at high exposure concentration. Cd exposure caused severe oxidative damage and cell necrosis in the tested bacterial strain. The co-exposure to PCP and Cd elevated the ROS level, stimulated the bacterial caspase activity, and induced DNA fragmentation, thereby leading to an apoptosis-like cell death. In conclusion, PCP-Cd complex pollution can cause bacterial population to decrease through apoptosis-like cell death pathway. However, it is worth noting that the subpopulation survives under the complex pollution stress.

Responses of bacterial communities along vertical soil profile to the chromium-contamination stress

Driven by rainfall and agricultural irrigation, chromium slag causes serious pollution to the subsurface environment and damages the indigenous soil bacterial community. The study of the ecological responses of bacterial community to chromium pollution can provide support for bioremediation of chromium pollution. In this study, soil samples at different depths (0, 2, 5, 7 m) in a long-term chromium contaminated site were selected to analyze the vertical changes and responses of bacterial community structure using high-throughput 16S rRNA gene sequencing. Total Cr and soluble Cr(VI) concentrations were highest in topsoil and decreased with depth. Compared with the topsoil of uncontaminated land, the alpha-diversity in the contaminated topsoil was significantly reduced, and the bacterial community composition was altered. With increasing soil depth, the alpha-diversity decreased, and the species distribution became more uneven. Topsoil and subsurface soil showed different bacterial community structures, associated with the Cr concentration and physicochemical properties. Proteobacteria in all samples were dominant, and their relative abundances increased with depth. Bacterial community alpha-diversity and dominant taxa can be used as indicators of response to chromium pollution. The predicted metagenomic functional annotation (using Tax4Fun) showed that the relative abundance of chromate transporters (chrA, K07240) in the topsoil is 20–30 times of that in deeper soil, indicating potential bioremediation capability by topsoil bacterial community. The bioremediation feasibility of reducing Cr(VI) using indigenous bacterial community showed that Cr-polluted topsoil bacterial community has ability to reduce Cr(VI).

Plant Growth Promoting Bacterial Consortium Induces Shifts in Indigenous Soil Bacterial Communities and Controls Listeria monocytogenes in Rhizospheres of Cajanus cajan and Festuca arundinacea.

The rhizosphere is a dynamic and complex interface between plant roots and microorganisms. Owing to exudates, a web of interactions establishes among the microbial members of this micro-environment. The present study explored the impact of a bacterial consortium (Azotobacter chroococcum, Bacillus megaterium and Pseudomonas fluorescens, ABP), on the fate of a human pathogen, Listeria monocytogenes EGD-e, in soil and in the rhizospheres of Cajanus cajan and Festuca arundinacea, in addition to its plant growth promoting effect. The study further assessed the impact these bioinoculants exert on the autochthonous soil bacterial communities. Experiments in sterilised soil inoculated with bioinoculants and L. monocytogenes revealed the inhibition of L. monocytogenes by approximately 80-fold compared to that without the consortium. Subsequently, experiments were conducted in non-sterile soil microcosms planted with C. cajan and F. arundinacea, and in bulk soil. The consortium led to a significant increase in plant growth in both plants and prevented growth of L. monocytogenes. However, the presence of resident soil bacterial communities overshadowed this inhibitory effect, and a sharp decline in L. monocytogenes populations (5-6 log reduction) was recorded under non-sterile soil conditions. A shift in the soil resident bacterial communities was observed upon amendment with the bioinoculants. A significant increase of potential Plant Growth Promoting Rhizobacteria (PGPR) and biocontrol agents was observed, while the abundance of potential phytopathogens dropped. The present study opens up new avenues for the application of such a consortium given their dual benefits of plant growth promotion and restricting phytopathogens as well as human pathogen.

Effect of protists on rhizobacterial community composition and rice plant growth in a biochar amended soil

The effects of protists on an indigenous soil bacterial community, putative bacterial genes involved in N-cycling, and the rice plant growth were studied in poultry litter biochar (PL) and rice husk biochar (RH) amended (with two application doses: 2% and 4% w/w) paddy field soil. The bacterial community composition, which was evaluated using 16S rRNA gene amplicon sequencing, was significantly and differentially affected by the protists, the PL and the RH. The effects of protists on the bacterial community composition were decreased by the RH and the PL treatments. The number of protist-affected bioindicator bacterial taxa was decreased from 90 to 46, 29, 43, and 21 in the 2% RH-, 4% RH-, 2% PL-, and 4% PL-treated soils, respectively. The presence of the protist significantly increased the abundance of the putative bacterial genes involved in mineralisation, dissimilatory nitrate reduction to ammonium (DNRA), and NO3- assimilation, and the same occurred with PL treatments. The rice plant growth and N uptake were always higher in the presence of protists and PL amendments. Overall our results suggest a new insight into the effects of biochar on the bacterial community via altering the trophic interactions.

Bacteriophages from Arsenic-Resistant Bacteria Transduced Resistance Genes, which Changed Arsenic Speciation and Increased Soil Toxicity

Lysogenic phages are known to serve as transfer vectors for bacterial genes involved in biotransformation of various environmental pollutants. However, their role in arsenic-contaminated environments is largely undocumented. Here, lysogenic phages were chemically induced (with mitomycin C) in soil samples from two contaminated sites, and arsenic resistance genes arsC (coding for As(V) reduction to excretable (via efflux pumps) but more toxic As(III)) and arsM (coding for As(III) methylation) were detected in these phage genomes. The relative abundance of these genes (per phage particle) was positively correlated with that in the corresponding indigenous soil bacterial communities (resistance gene per 16S rRNA), with R2 = 0.974 for arsC and 0.761 for arsM. Microcosm studies with 100 mg/kg of arsenic soil showed that phages (amended at 5.0 × 107 phages per gram soil) enhanced the propagation of arsC by 122-fold and arsM by 575-fold, relative to unamended soil. This increased the As(III) concentration by 4.3...

Sequential biowashing-biopile processes for remediation of crude oil contaminated soil in Kuwait.

The application of biological processes for remediation of the aged crude oil-contaminated soil of Kuwait can be an inefficient way, thus, this study developed 20 d-sequential biowashing and biopile processes where the biowashing step uses an enrichment culture of the indigenous soil bacterial community and the biopile step includes hemoglobin-catalyzed oxidation (HCO). The residual total petroleum hydrocarbons (TPH) concentrations and CO2 generation were measured to determine the removal efficiency, and the bacterial community changes were studied to investigate the effect of the sequential processes on the soil indigenous bacterial community. The enrichment culture grown on hemoglobin showed an increased surface activity, and this promoted desorption and emulsification of crude oil from the soil sample in the biowashing step resulting in 75% TPH removal. Potential surfactant-producing bacterial species were observed in the soil sample after biowashing. The HCO in the beginning of the biopile step removed 21% of the residual TPH, and further TPH removal was observed with a longer biopile period. Overall, the sequential biowashing and biopile processes removed 86% TPH. The results show that the developed sequential biowashing and biopile processes can be used to efficiently remediate the aged crude oil-contaminated soil of Kuwait.

Traits-Based Integration of Multi-Species Inoculants Facilitates Shifts of Indigenous Soil Bacterial Community.

Microbial co-inoculation is considered to be an innovative approach and had been applied worldwide. However, the underlying mechanisms of microbial co-inoculants constructions, especially the trait-based combination of distinctly different microbial species remains poorly understood. In this study, we constructed two microbial co-inoculants with the same three strains with emphasis on the microbial, soil and plant traits. Microbial co-inoculants 1 (M1) were constructed according to soil fertility, microbial activity and cucumber nutrient requirement with a 2:1:2 ratio (Ensifer sp. NYM3, Acinetobacter sp. P16 and Flavobacterium sp. KYM3), while microbial co-inoculants 2 (M2) were constructed according to soil fertility and cucumber nutrient requirement with a 1:10:1 ratio without considering the difference in the nutrient supply capability of microbial species. The results showed that M1 and M2 both obviously increased cucumber yields. The M1 had significant highest pH value, total nitrogen (TN) and invertase activity (IA). The M2 had significant highest available phosphate (AP), NO3-N, urea activity (UA), and alkaline phosphatase activity (APA). Gammaproteobacteria, Acidobacteria, Nitrospirae, and Armatimonadetes were significantly increased, while Actinobacteria and Firmicutes were significantly decreased by microbial co-inoculations (M1 and M2). The bacterial lineages enriched in M1 were Gammaproteobacteria and TM7. Acidobacteria, Bacteroidetes, and Deltaproteobacteria were enriched in M2. Principal coordinate analysis (PCoA) analysis showed that the bacterial communities were strongly separated by the different microbial inoculation treatments. The functional groups of intracellular_parasites were highest in M1. The functional groups of phototrophy, photoautotrophy, nitrification, fermentation, cyanobacteria, oxygenic_photoautotrophy, chitinolysis and animal_parasites_or_symbionts were highest in M2. Based on correlation analysis, it inferred that the M1 and M2 might promote cucumber yields by mediating bacterial community structure and function about nitrogen fixing and urea-N hydrolysis, respectively. Collectively, these results revealed that microbial co-inoculants had positive effects on cucumber yields. Trait-based integration of different microbial species had significant effects on soil properties and bacterial communities. It indicated that microbial activity should be considered in the construction of microbial co-inoculants. This will expand our knowledge in bacteria interaction, deepen understanding of microbial inoculants in improving plant performance, and will guide microbial fertilizer formulation and application in future.

Bioaugmentation of thiabendazole-contaminated soils from a wastewater disposal site: Factors driving the efficacy of this strategy and the diversity of the indigenous soil bacterial community.

The application of the fungicide thiabendazole (TBZ) in fruit packaging plants (FPP) results in the production of effluents which are often disposed in adjacent field sites. These require remediation to prevent further environmental dispersal of TBZ. We assessed the bioaugmentation potential of a newly isolated TBZ-degrading bacterial consortium in a naturally contaminated soil (NCS) exhibiting a natural gradient of TBZ levels (12000, 400, 250 and 12mgkg-1). The effect of aging on bioaugmentation efficacy was comparatively tested in a soil with similar physicochemical properties and soil microbiota, which was artificially, contaminated with the same TBZ levels (ACS). The impact of bioaugmentation and TBZ on the bacterial diversity in the NCS was explored via amplicon sequencing. Bioaugmentation effectively removed TBZ from both soils at levels up to 400mgkg-1 but failed at the highest contamination level (12000mgkg-1). Dissipation of TBZ in bioaugmented samples showed a concentration-dependent pattern, while aging of TBZ had a slight effect on bioaugmentation efficiency. Bioaugmentation had no impact on the soil bacterial diversity, in contrast to TBZ contamination. Soils from the hotspots of TBZ contamination (12000mgkg-1) showed a drastically lower α-diversity driven by the dominance of β- and γ-proteobacteria at the expense of all other bacterial phyla, especially Actinobacteria. Overall, bioaugmentation with specialized microbial inocula could be an effective solution for the recovery of disposal sites contaminated with persistent chemicals like TBZ.

Daily thanatomicrobiome changes in soil as an approach of postmortem interval estimation: An ecological perspective

Understanding human decomposition is critical for its use in postmortem interval (PMI) estimation, having a significant impact on forensic investigations. In recognition of the need to establish the scientific basis for PMI estimation, several studies on decomposition have been carried out in the last years. The aims of the present study were: (i) to identify soil microbiota communities involved in human decomposition through high-throughput sequencing (HTS) of DNA sequences from the different bacteria, (ii) to monitor quantitatively and qualitatively the decay of such signature species, and (iii) to describe succesional changes in bacterial populations from the early putrefaction state until skeletonization. Three donated individuals to the University of Tennessee FAC were studied. Soil samples around the body were taken from the placement of the donor until advanced decay/dry remains stage. Bacterial DNA extracts were obtained from the samples, HTS techniques were applied and bioinformatic data analysis was performed. The three cadavers showed similar overall successional changes. At the beginning of the decomposition process the soil microbiome consisted of diverse indigenous soil bacterial communities. As decomposition advanced, Firmicutes community abundance increased in the soil during the bloat stage. The growth curve of Firmicutes from human remains can be used to estimate time since death during Tennessee summer conditions.

Alterations in soil microbial communities caused by treatments with penicillin or neomycin

Antibiotic residues in soils can lead to serious health risk and ecological hazards. In this study, the effects of penicillin and neomycin, two antibiotics widely used in animal production, were investigated on soil bacterial communities. Changes in the community structure were monitored using three 16S ribosomal DNA (rDNA) polymerase chain reaction-based approaches, including denaturing gradient gel electrophoresis (DGGE), amplified rDNA restriction analysis (ARDRA), and terminal-restriction fragment length polymorphism (T-RFLP) analysis. The prominent DGGE bands were excised from gels and sequenced, and the data indicated the prevalence of Gammaproteobacteria in the soils. The total soil bacterial community, including uncultured bacteria, exhibited a higher diversity than that of cultured bacteria. Some microbial strains were capable of surviving and even subsisting on penicillin or neomycin. We also observed toxic effects of the antibiotics on the indigenous soil bacterial communities since some genotypes disappeared after the treatments (e.g., Pseudomonas sp., Stenotrophomonas sp., Salinimonas, and uncultured Acinetobacter sp.). The implications of these findings are that the functions of soil bacterial communities may be negatively affected if key microbial community members are lost.

상수리나무림의 토양 층위별 세균군집의 계통학적 특성

Department of Microbial & Nanomaterials, Mokwon University, Daejeon 302-729, Republic of Korea(Received April 24, 2015; Accepted June 19, 2015)ABSTRACT: We have examined the correlation between the physicochemical and microbiological environment variables for the different layers of oak forest soil in Mt. Gyeryong, Korea. The result shows that there is a high correlation in the environment variable s between the soil parameters of the fermented (F) layer and humus (H) layer. In particular, the pH level in the F layer shows a high correla tion with C and N, while the various organic acids of the H layer turns out to be closely correlated with soil bacteria density. As we evaluated phylogenetic characteristics of bacterial populations by DGGE analysis with DNA extracted. Total of 175 bands including 43 band s from litter (L) layer, 42 bands from F layer, 43 bands from H layer and 47 bands from rhizosphere (A) layer were selected as the maj or DGGE band of oak forest soil. Based on the 16S rRNA gene sequences, 175 DGGE bands were classified into 32 orders in 7 phylum. The heat map was analyzed in order to compare the quantity of the base sequences of each order and based on the clustering of the different layers of oak forest soil, the result confirms that the F layer and H layer belong to a different cluster from that of L layer and A layer. Furthermore, it also showed that approximately 50% of the total microbial population in different layers is α-proteobacteria , which indicates that they belong to the dominant system group. In particular, Rhizobiales , Burkholderiales and Actinobacteriales were observed in all the seasons and layers of oak forest soil, which confirms that they are the indigenous soil bacterial community in oak forest soil.Key words: bacterial populations, DGGE, oak forest soil, phylogeny

Assessing Long Term Effects of Bioremediation: Soil Bacterial Communities 14 Years after Polycyclic Aromatic Hydrocarbon Contamination and Introduction of a Genetically Engineered Microorganism

Environmental contamination by organics such as polycyclic aromatic hydrocarbons (PAHs) generally changes native microbial communities. However our understanding of microbial responses has been limited to short term studies (i.e., less than 2-3 years) so long term community responses are not as well understood. In 1996, the genetically engineered microorganism Pseudomonas fluorescens HK44 was released into polycyclic aromatic hydrocarbon (PAH)-contaminated soil in lysimeters to monitor in situ PAH-biodegradation. The objective of this study was to assess the long term impacts of PAH contamination and addition of HK44 on the indigenous soil bacterial community structure. In 2010, 14 years after the lysimeter experiment initiation, lysimeters were unsealed and sampled. Although PAHs were degraded and PAH concentrations fell below detectable levels within approximately the first two years of this experiment, lysimeters that had received PAHs had significantly higher soil organic matter content (1.30 ± 0.23%) than control lysimeters with clean soils (0.81 ± 0.08%). Pyrosequencing of 16S rRNA gene amplicon libraries revealed a distinct bacterial community structure in the lysimeters that had received PAHs. In contrast, there were no discernible differences in soil chemistry or bacterial community structures in lysimeters where HK44 was inoculated compared to those to which HK44 was not inoculated. These results indicate that although the initial perturbations are no longer detectable, the addition of PAHs had long term influences on the bacterial communities, while the introduction of the genetically engineered microorganism HK44 did not.

Pyrosequence analyses of bacterial communities during simulated in situ bioremediation of polycyclic aromatic hydrocarbon-contaminated soil.

Barcoded amplicon pyrosequencing was used to generate libraries of partial 16S rRNA genes from two columns designed to simulate in situ bioremediation of polycyclic aromatic hydrocarbons (PAHs) in weathered, contaminated soil. Both columns received a continuous flow of artificial groundwater but one of the columns additionally tested the impact of biostimulation with oxygen and inorganic nutrients on indigenous soil bacterial communities. The penetration of oxygen to previously anoxic regions of the columns resulted in the most significant community changes. PAH-degrading bacteria previously determined by stable-isotope probing (SIP) of the untreated soil generally responded negatively to the treatment conditions, with only members of the Acidovorax and a group of uncharacterized PAH-degrading Gammaproteobacteria maintaining a significant presence in the columns. Additional groups of sequences associated with the Betaproteobacterial family Rhodocyclaceae (including those associated with PAH degradation in other soils), and the Thiobacillus, Thermomonas, and Bradyrhizobium genera were also present in high abundance in the biostimulated column. Similar community responses were previously observed during biostimulated ex situ treatment of the same soil in aerobic, slurry-phase bioreactors. While the low relative abundance of many SIP-determined groups in the column libraries may be a reflection of the slow removal of PAHs in that system, the similar response of known PAH degraders in a higher-rate bioreactor system suggests that alternative PAH-degrading bacteria, unidentified by SIP of the untreated soil, may also be enriched in engineered systems.

Compositional shifts of bacterial groups in a solarized and amended soil as determined by denaturing gradient gel electrophoresis

Soil solarization is a widespread, nonchemical agricultural practice for disinfesting soils, which is often used in combination with organic amendment, and whose action represents an important factor impacting on soil bacterial communities structure and population dynamics. The present study was conducted to investigate whether and to which extent a 72-day plot-scale soil solarization treatment, either combined or not with organic amendment, could stimulate compositional changes in the genetic structure of indigenous soil bacterial communities. Soil solarization with transparent polyethylene film, in combination or not with farmyard manure addition, was carried out during a summer period on a clay loam agricultural soil located in Southern Italy. Soils from a four-treatment (NS, nonsolarized control soil; S, solarized soil; MA, manure-amended nonsolarized soil; MS, manure-amended and solarized soil) plot block were sampled after 0, 8, 16, 36 and 72 days. Compositional shifts in the genetic structure of indigenous soil bacterial communities were monitored by denaturing gradient gel electrophoresis (DGGE) fingerprinting of 16S rRNA gene fragments amplified from soil-extracted community DNA using primers specific for Bacteria, Actinomycetales, α- and β-Proteobacteria. Changes in soil temperature, pH, and electrical conductivity (EC 1:1) were also monitored from 0 to 72 days. Beneath the polyethylene film the average soil temperature at 8-cm depth reached 55 °C compared to 35 °C in nonsolarized soil. In general, without amendment both soil pH and EC 1:1 were not significantly affected by solarization, whereas in manured plots either variables were greatly increased (from 7.0 to 8.0 pH and from 271 to 3021 μS cm −1 EC 1:1), and both showed long-lasting effects due to soil solar heating. The eubacterial DGGE profiles revealed that soil solarization was the main factor inducing strong time-dependent population shifts in the community structure either in unamended or amended soils. Conversely, the addition of organic amendment resulted in an altered bacterial community, which remained rather stable over time. A similar behaviour was also observed in the DGGE patterns of β-proteobacterial and actinomycete populations, and also, albeit to a lesser extent, in the DGGE profiles of α-Proteobacteria. An increased bacterial richness was evidenced by DGGE fingerprints in 16- and 36-day samplings, followed by a decrease appearing in 72-day samplings. This could be explained, other than by a direct thermal effect on soil microflora, by solarization-induced changes in the physico-chemical properties of soil microbial habitats or by other ecological factors (e.g. decreased competitiveness of dominating bacterial species, reduced grazing pressure of microfaunal predators, increased nutrient availability).

Impact of DNA extraction method on bacterial community composition measured by denaturing gradient gel electrophoresis

The impact of DNA extraction protocol on soil DNA yield and bacterial community composition was evaluated. Three different procedures to physically disrupt cells were compared: sonication, grinding–freezing–thawing, and bead beating. The three protocols were applied to three different topsoils. For all soils, we found that each DNA extraction method resulted in unique community patterns as measured by denaturing gradient gel electrophoresis. This indicates the importance of the DNA extraction protocol on data for evaluating soil bacterial diversity. Consistently, the bead-beating procedure gave rise to the highest number of DNA bands, indicating the highest number of bacterial species. Supplementing the bead-beating procedure with additional cell-rupture steps generally did not change the bacterial community profile. The same consistency was not observed when evaluating the efficiency of the different methods on soil DNA yield. This parameter depended on soil type. The DNA size was of highest molecular weight with the sonication and grinding–freezing–thawing procedures (approx. 20 kb). In contrast, the inclusion of bead beating resulted in more sheared DNA (approx. 6–20 kb), and the longer the bead-beating time, the higher the fraction of low-molecular weight DNA. Clearly, the choice of DNA extraction protocol depends on soil type. We found, however, that for the analysis of indigenous soil bacterial communities the bead-beating procedure was appropriate because it is fast, reproducible, and gives very pure DNA of relatively high molecular weight. And very importantly, with this protocol the highest soil bacterial diversity was obtained. We believe that the choice of DNA extraction protocol will influence not only the determined phylogenetic diversity of indigenous microbial communities, but also the obtained functional diversity. This means that the detected presence of a functional gene—and thus the indication of enzyme activity—may depend on the nature of the applied DNA extraction procedure.

Effects of compost addition and simulated solarisation on the fate of Ralstonia solanacearum biovar 2 and indigenous bacteria in soil

Abstract The effects of compost addition and simulated solarisation of soil on the survival of Ralstonia solanacearum biovar 2 strain 1609, as well as on the structure of indigenous soil bacterial communities, were analysed. In addition, effects on the invasion of susceptible test plants by strain 1609 were assessed. In untreated soil in microcosms and the field, strain 1609 showed slow progressive declines, from 10(6)-10(7) to roughly 10(4)-10(5) CFU per g dry soil in around 60 days. When these soils were used in suppressiveness tests, a majority of plants developed symptoms of wilting and revealed the presence of the pathogen in their lower stem parts, as evidenced by immunofluorescence colony staining (IFC) and polymerase chain reaction (PCR). Solarisation of unamended soil did not drastically affect R. solanacearum survival or plant invasiveness. However, the addition of household compost resulted in enhanced R. solanacearum population decline rates, as well as reduced numbers of diseased plants in suppressiveness tests. Combined solarisation and compost addition yielded differential results between microcosms and the field. Some healthy-looking plants, primarily from soils treated with compost, revealed the latent presence of strain 1609 in the lower stem parts. The eubacterial and beta-subgroup proteobacterial communities in the differentially treated soil microcosms were rather stable, as evidenced by analysis of PCR-denaturing gradient gel electrophoresis (DGGE) generated molecular profiles. However, compost amendment clearly induced changes in these communities, which were detectable until the end of the experiment; two major bands, affiliated with Variovorax paradoxus and Aquaspirillum psychrophylum, were associated with the compost amendment. The decrease in abundance of R. solanacearum in the compost-amended soils was confirmed by the DGGE profiles.

Effect of experimental contamination with the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine on soil bacterial communities

The effect of contamination with the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) on an indigenous soil bacterial community was examined in two uncontaminated loam soil columns possessing native grasses. One column was spiked twice with RDX crystals for a total RDX load of 1000 mg (kg soil) −1. The reduced metabolite of RDX degradation, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine, was observed in the column leachate, suggesting anaerobic degradation of RDX. Denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA from both contaminated and uncontaminated columns produced identical banding patterns which were stable over the course of the experimental period. The bacterial diversity remained high in the contaminated column, as determined by restriction fragment length polymorphism and rarefaction analyses of random 16S rDNA clones. These combined results suggested that long-term exposure to 1000 mg RDX (kg soil) −1 did not produce an observable effect on bacterial diversity or the numerically dominant members of the indigenous soil bacterial community.

Effect of experimental contamination with the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine on soil bacterial communities

The effect of contamination with the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) on an indigenous soil bacterial community was examined in two uncontaminated loam soil columns possessing native grasses. One column was spiked twice with RDX crystals for a total RDX load of 1000 mg (kg soil)(-1). The reduced metabolite of RDX degradation, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine, was observed in the column leachate, suggesting anaerobic degradation of RDX. Denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA from both contaminated and uncontaminated columns produced identical banding patterns which were stable over the course of the experimental period. The bacterial diversity remained high in the contaminated column, as determined by restriction fragment length polymorphism and rarefaction analyses of random 16S rDNA clones. These combined results suggested that long-term exposure to 1000 mg RDX (kg soil)(-1) did not produce an observable effect on bacterial diversity or the numerically dominant members of the indigenous soil bacterial community.

Effects of compost addition and simulated solarisation on the fate of Ralstonia solanacearum biovar 2 and indigenous bacteria in soil

The effects of compost addition and simulated solarisation of soil on the survival of Ralstonia solanacearum biovar 2 strain 1609, as well as on the structure of indigenous soil bacterial communities, were analysed. In addition, effects on the invasion of susceptible test plants by strain 1609 were assessed. In untreated soil in microcosms and the field, strain 1609 showed slow progressive declines, from 10 6–10 7 to roughly 10 4–10 5 CFU per g dry soil in around 60 days. When these soils were used in suppressiveness tests, a majority of plants developed symptoms of wilting and revealed the presence of the pathogen in their lower stem parts, as evidenced by immunofluorescence colony staining (IFC) and polymerase chain reaction (PCR). Solarisation of unamended soil did not drastically affect R. solanacearum survival or plant invasiveness. However, the addition of household compost resulted in enhanced R. solanacearum population decline rates, as well as reduced numbers of diseased plants in suppressiveness tests. Combined solarisation and compost addition yielded differential results between microcosms and the field. Some healthy-looking plants, primarily from soils treated with compost, revealed the latent presence of strain 1609 in the lower stem parts. The eubacterial and β-subgroup proteobacterial communities in the differentially treated soil microcosms were rather stable, as evidenced by analysis of PCR-denaturing gradient gel electrophoresis (DGGE) generated molecular profiles. However, compost amendment clearly induced changes in these communities, which were detectable until the end of the experiment; two major bands, affiliated with Variovorax paradoxus and Aquaspirillum psychrophylum, were associated with the compost amendment. The decrease in abundance of R. solanacearum in the compost-amended soils was confirmed by the DGGE profiles.